Contribution of extracellular vesicles (EV) in epithelial-mesenchymal transition
The metastatic cascade describes the process by which tumour cells escape their primary site and colonize secondary locations. Tumour angiogenesis facilitates passage, and cells at the leading edge of the primary tumour are thought to undergo epithelial-mesenchymal transition (EMT) to acquire increased motility and invasiveness. Whether oncogenic cells that have undergone EMT directly promotes endothelial cell recruitment remains largely unknown, and the role of extracellular vesicles (EVs) (30-1,000nm diameter) in this process has not yet been definitively explored.
Conventional biological assays for cell proliferation, motility, migration, and invasion are already established in our lab. Using global profiling approaches, including proteomic (mass-spectrometry based profiling) and genomic (miR/mRNA profiling) analyses, we intend to catalogue and identify the contribution of EVs during EMT, especially induced expression of signalling pathway receptors/ modulators in recipient cells to modify their function.
Preliminary results demonstrate that oncogenic cells undergoing EMT can communicate with endothelial cells via exosomes, and establish angiogenic promoters that may function during the initial stages of metastasis.
This project will investigate the following questions:
(1) What are the different methods to purify the secretome and distinct EV subtypes?
(2) Can distinct cargo components in EVs be perturbed using molecular biology?
(3) Investigate the contribution of secreted (soluble and vesicle components) to modifying recipient cell function.
(4) How do vesicles selectively package and transfer soluble cargo such as cellular mediators directly into endothelial and epithelial cells?